the biological activity of certain derivatives of ddt
TRANSCRIPT
THE BIOLOGICAL ACTIVITY OF CERTAIN DERIVATIVES OF DM
by
JO~EPH ELI MOORE
A THESIS
submitted to
OREGON STATE COLLEGE
1n partial fulfillment of the requirements for the
degree of
DOCTOR OF PHILOSOPHY
June 1954
APFBOTEDT
Redacted for Privacy
Asltrtmt Prsfrsscr sf Ghamtstry
h Chergc of te tor
Redacted for Privacyaf Drpartncnt cf Chenlstry
Redacted for PrivacyChrtmn cf $chacl Orrdurtc Ootrnltt
Redacted for PrivacyDma cf Gmdurta $obsat
Dltr lfhorle 1r prorcntrd
$rgd bf bfllte foarc
ACtNOWLEDGMENT
The author wishes to express sincere thanks to
Dr Edward c Bubl for his understanding and experienced
advice during h1s supervision ot this thesis
Credit must also be given to Dr Bubl for developshy
ment of the gradient plate technique and to Dr teon c Terriere and MJt David w Ingalsbe who kindly perfoimed
the screening of compounds on mosquito larvae
TABLE 0 F CONT NTS
Introdu~tion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull
Experimental bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Synthesis of Compounds bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Compound Screening on Mosquito Larvae bullbullbullbull 21
Compound Screening on Microorganisms bullbullbullbullbull 22
F1gur 1 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23middot
Figure 2 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23
Figure 3 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 24
Figure 4 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 25
Table I bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 26
Sample Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 27
Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 28
Discussion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 33
Sununary bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 36
Bibliography ~ bullbullbullbullbull ~ bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull 37
Addendum bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bullbull bull 42
Table III bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 45
Bibliography bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 47
THE BIOWGICAL ACTIVITY OF CERTAIN DERIVATIVES OF DDT
INTRODUCTION
It has long been recognized that the discovery of
a law rolating biological activity of organic compounds
to their structures would be of benefit to the investishy
gator who is unceasingly searching for agents exhibiting
such activity Selectively toxc agents in particular
are needed~ for agricultural foodstuffs must be protected
from weeds and pests of many kinds and economic animals
are troUbled with biological enemies of all types an
himself is n~t respected by these parasites
Up to this period of development in various fields
of the sciences ttempts have been made to correlate
chemical structure wit~ specific selective toxic activity
The insecticide DDT ll-b1s-(p-chlorophenyl)222-tribull
chloroethane m y be taken as a prime exam le for because
of its great economic importance many investigators have
given their time to considering this biologically active
compound
A first group considers the toxio portion of the
DDT molecule as res1d1ng 1n the linked p-ohlorobenzene
rings while the -cc13 grouping is believed to impart lipshy
oid solubility (22 pp892-928) Another group suggests
2
that the linked pbulloblorobenzene rings impart lipoid solushy
bility while toxioitr is due rather to the bility to split
out hydrogen chloride from the remainder of the molecule
(25 pp51~-513) A third group casts doubt on either of
these speculations and oonsidels that perhaps too simple a
relationship is sought tor such a complex phenomenon (6
ppl69-170)
A more theoretical approach has been taken recently
by yet anothe~ group who feels there is little basis for
assigning more or less independent lipoid solubility and
toxic properties to parts of the DDT molecule (38 pp2990shy
2999) Rather they consider that toxicity is due to the
molecule as a whole and is explainable by means of a tr1shy
hedral1zed structure resulting from interaction of the
p-subst1tuted benzene rings and the bullCC13 group or groupshy
ings of similar and greater size
However a la relating toxicity to structure
broad enough to systematize and sympllfy the development
of successful agents needed t~ combat parasites afflicting
Man and his f~~d sources has not been forthcoming
The first step in any logical approach as to how
bigtlog1cally active substances w~rk is to record the
various biological physical and chemical properties of
active compounds~ Until enough relevant facts are colshy
lected progress toward such an end is slow and time
consuming Therefore it 1s hoped that the data g1ven
herein will add a small part to the necessary accumulation
ot evidence that may eventually settle the question of this
correlation or lack of correlation between chemical strucshy
ture and toxicity
The d1phenylmethylene radical Q bullCbull Q 1 was
chosen for this study of structural relationship to toxicshy
ity A review of the literature shows that this molecular
arrangement is contained in a number of biologically active
compounds
A preliminary investigation was first made in which
several derivatives of the basic structure were synthesized
in order to ascertain 1f certain functional groups ould
most readily lend themselvemiddota to study The results of th18
preliminary investigation indicated th t additional time
might pr~titably be spent contrasting the biological activshy
ity of ketone der-ivatives with the corresponding alcohols
Accordingly there were synthesized a series of
ketmes secondary alcohols and in the cases found feasible
the tertiary alcohols bull Chemical and biological data on
these compounds are given herein Other derivatives studied
in the preliminary investigation are als~ listed in an
addendum
The compounds were screened on mosquito larvae and
six microorganisms chosen for this purpose
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
APFBOTEDT
Redacted for Privacy
Asltrtmt Prsfrsscr sf Ghamtstry
h Chergc of te tor
Redacted for Privacyaf Drpartncnt cf Chenlstry
Redacted for PrivacyChrtmn cf $chacl Orrdurtc Ootrnltt
Redacted for PrivacyDma cf Gmdurta $obsat
Dltr lfhorle 1r prorcntrd
$rgd bf bfllte foarc
ACtNOWLEDGMENT
The author wishes to express sincere thanks to
Dr Edward c Bubl for his understanding and experienced
advice during h1s supervision ot this thesis
Credit must also be given to Dr Bubl for developshy
ment of the gradient plate technique and to Dr teon c Terriere and MJt David w Ingalsbe who kindly perfoimed
the screening of compounds on mosquito larvae
TABLE 0 F CONT NTS
Introdu~tion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull
Experimental bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Synthesis of Compounds bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Compound Screening on Mosquito Larvae bullbullbullbull 21
Compound Screening on Microorganisms bullbullbullbullbull 22
F1gur 1 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23middot
Figure 2 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23
Figure 3 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 24
Figure 4 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 25
Table I bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 26
Sample Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 27
Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 28
Discussion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 33
Sununary bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 36
Bibliography ~ bullbullbullbullbull ~ bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull 37
Addendum bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bullbull bull 42
Table III bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 45
Bibliography bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 47
THE BIOWGICAL ACTIVITY OF CERTAIN DERIVATIVES OF DDT
INTRODUCTION
It has long been recognized that the discovery of
a law rolating biological activity of organic compounds
to their structures would be of benefit to the investishy
gator who is unceasingly searching for agents exhibiting
such activity Selectively toxc agents in particular
are needed~ for agricultural foodstuffs must be protected
from weeds and pests of many kinds and economic animals
are troUbled with biological enemies of all types an
himself is n~t respected by these parasites
Up to this period of development in various fields
of the sciences ttempts have been made to correlate
chemical structure wit~ specific selective toxic activity
The insecticide DDT ll-b1s-(p-chlorophenyl)222-tribull
chloroethane m y be taken as a prime exam le for because
of its great economic importance many investigators have
given their time to considering this biologically active
compound
A first group considers the toxio portion of the
DDT molecule as res1d1ng 1n the linked p-ohlorobenzene
rings while the -cc13 grouping is believed to impart lipshy
oid solubility (22 pp892-928) Another group suggests
2
that the linked pbulloblorobenzene rings impart lipoid solushy
bility while toxioitr is due rather to the bility to split
out hydrogen chloride from the remainder of the molecule
(25 pp51~-513) A third group casts doubt on either of
these speculations and oonsidels that perhaps too simple a
relationship is sought tor such a complex phenomenon (6
ppl69-170)
A more theoretical approach has been taken recently
by yet anothe~ group who feels there is little basis for
assigning more or less independent lipoid solubility and
toxic properties to parts of the DDT molecule (38 pp2990shy
2999) Rather they consider that toxicity is due to the
molecule as a whole and is explainable by means of a tr1shy
hedral1zed structure resulting from interaction of the
p-subst1tuted benzene rings and the bullCC13 group or groupshy
ings of similar and greater size
However a la relating toxicity to structure
broad enough to systematize and sympllfy the development
of successful agents needed t~ combat parasites afflicting
Man and his f~~d sources has not been forthcoming
The first step in any logical approach as to how
bigtlog1cally active substances w~rk is to record the
various biological physical and chemical properties of
active compounds~ Until enough relevant facts are colshy
lected progress toward such an end is slow and time
consuming Therefore it 1s hoped that the data g1ven
herein will add a small part to the necessary accumulation
ot evidence that may eventually settle the question of this
correlation or lack of correlation between chemical strucshy
ture and toxicity
The d1phenylmethylene radical Q bullCbull Q 1 was
chosen for this study of structural relationship to toxicshy
ity A review of the literature shows that this molecular
arrangement is contained in a number of biologically active
compounds
A preliminary investigation was first made in which
several derivatives of the basic structure were synthesized
in order to ascertain 1f certain functional groups ould
most readily lend themselvemiddota to study The results of th18
preliminary investigation indicated th t additional time
might pr~titably be spent contrasting the biological activshy
ity of ketone der-ivatives with the corresponding alcohols
Accordingly there were synthesized a series of
ketmes secondary alcohols and in the cases found feasible
the tertiary alcohols bull Chemical and biological data on
these compounds are given herein Other derivatives studied
in the preliminary investigation are als~ listed in an
addendum
The compounds were screened on mosquito larvae and
six microorganisms chosen for this purpose
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
ACtNOWLEDGMENT
The author wishes to express sincere thanks to
Dr Edward c Bubl for his understanding and experienced
advice during h1s supervision ot this thesis
Credit must also be given to Dr Bubl for developshy
ment of the gradient plate technique and to Dr teon c Terriere and MJt David w Ingalsbe who kindly perfoimed
the screening of compounds on mosquito larvae
TABLE 0 F CONT NTS
Introdu~tion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull
Experimental bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Synthesis of Compounds bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Compound Screening on Mosquito Larvae bullbullbullbull 21
Compound Screening on Microorganisms bullbullbullbullbull 22
F1gur 1 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23middot
Figure 2 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23
Figure 3 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 24
Figure 4 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 25
Table I bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 26
Sample Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 27
Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 28
Discussion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 33
Sununary bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 36
Bibliography ~ bullbullbullbullbull ~ bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull 37
Addendum bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bullbull bull 42
Table III bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 45
Bibliography bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 47
THE BIOWGICAL ACTIVITY OF CERTAIN DERIVATIVES OF DDT
INTRODUCTION
It has long been recognized that the discovery of
a law rolating biological activity of organic compounds
to their structures would be of benefit to the investishy
gator who is unceasingly searching for agents exhibiting
such activity Selectively toxc agents in particular
are needed~ for agricultural foodstuffs must be protected
from weeds and pests of many kinds and economic animals
are troUbled with biological enemies of all types an
himself is n~t respected by these parasites
Up to this period of development in various fields
of the sciences ttempts have been made to correlate
chemical structure wit~ specific selective toxic activity
The insecticide DDT ll-b1s-(p-chlorophenyl)222-tribull
chloroethane m y be taken as a prime exam le for because
of its great economic importance many investigators have
given their time to considering this biologically active
compound
A first group considers the toxio portion of the
DDT molecule as res1d1ng 1n the linked p-ohlorobenzene
rings while the -cc13 grouping is believed to impart lipshy
oid solubility (22 pp892-928) Another group suggests
2
that the linked pbulloblorobenzene rings impart lipoid solushy
bility while toxioitr is due rather to the bility to split
out hydrogen chloride from the remainder of the molecule
(25 pp51~-513) A third group casts doubt on either of
these speculations and oonsidels that perhaps too simple a
relationship is sought tor such a complex phenomenon (6
ppl69-170)
A more theoretical approach has been taken recently
by yet anothe~ group who feels there is little basis for
assigning more or less independent lipoid solubility and
toxic properties to parts of the DDT molecule (38 pp2990shy
2999) Rather they consider that toxicity is due to the
molecule as a whole and is explainable by means of a tr1shy
hedral1zed structure resulting from interaction of the
p-subst1tuted benzene rings and the bullCC13 group or groupshy
ings of similar and greater size
However a la relating toxicity to structure
broad enough to systematize and sympllfy the development
of successful agents needed t~ combat parasites afflicting
Man and his f~~d sources has not been forthcoming
The first step in any logical approach as to how
bigtlog1cally active substances w~rk is to record the
various biological physical and chemical properties of
active compounds~ Until enough relevant facts are colshy
lected progress toward such an end is slow and time
consuming Therefore it 1s hoped that the data g1ven
herein will add a small part to the necessary accumulation
ot evidence that may eventually settle the question of this
correlation or lack of correlation between chemical strucshy
ture and toxicity
The d1phenylmethylene radical Q bullCbull Q 1 was
chosen for this study of structural relationship to toxicshy
ity A review of the literature shows that this molecular
arrangement is contained in a number of biologically active
compounds
A preliminary investigation was first made in which
several derivatives of the basic structure were synthesized
in order to ascertain 1f certain functional groups ould
most readily lend themselvemiddota to study The results of th18
preliminary investigation indicated th t additional time
might pr~titably be spent contrasting the biological activshy
ity of ketone der-ivatives with the corresponding alcohols
Accordingly there were synthesized a series of
ketmes secondary alcohols and in the cases found feasible
the tertiary alcohols bull Chemical and biological data on
these compounds are given herein Other derivatives studied
in the preliminary investigation are als~ listed in an
addendum
The compounds were screened on mosquito larvae and
six microorganisms chosen for this purpose
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
TABLE 0 F CONT NTS
Introdu~tion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull
Experimental bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Synthesis of Compounds bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 6
Compound Screening on Mosquito Larvae bullbullbullbull 21
Compound Screening on Microorganisms bullbullbullbullbull 22
F1gur 1 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23middot
Figure 2 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 23
Figure 3 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 24
Figure 4 bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 25
Table I bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 26
Sample Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 27
Table II bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 28
Discussion bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 33
Sununary bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 36
Bibliography ~ bullbullbullbullbull ~ bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull bull 37
Addendum bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bull bull bullbull bull bull bull bull bull bull bull bull bull bullbull bull 42
Table III bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 45
Bibliography bullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbullbull 47
THE BIOWGICAL ACTIVITY OF CERTAIN DERIVATIVES OF DDT
INTRODUCTION
It has long been recognized that the discovery of
a law rolating biological activity of organic compounds
to their structures would be of benefit to the investishy
gator who is unceasingly searching for agents exhibiting
such activity Selectively toxc agents in particular
are needed~ for agricultural foodstuffs must be protected
from weeds and pests of many kinds and economic animals
are troUbled with biological enemies of all types an
himself is n~t respected by these parasites
Up to this period of development in various fields
of the sciences ttempts have been made to correlate
chemical structure wit~ specific selective toxic activity
The insecticide DDT ll-b1s-(p-chlorophenyl)222-tribull
chloroethane m y be taken as a prime exam le for because
of its great economic importance many investigators have
given their time to considering this biologically active
compound
A first group considers the toxio portion of the
DDT molecule as res1d1ng 1n the linked p-ohlorobenzene
rings while the -cc13 grouping is believed to impart lipshy
oid solubility (22 pp892-928) Another group suggests
2
that the linked pbulloblorobenzene rings impart lipoid solushy
bility while toxioitr is due rather to the bility to split
out hydrogen chloride from the remainder of the molecule
(25 pp51~-513) A third group casts doubt on either of
these speculations and oonsidels that perhaps too simple a
relationship is sought tor such a complex phenomenon (6
ppl69-170)
A more theoretical approach has been taken recently
by yet anothe~ group who feels there is little basis for
assigning more or less independent lipoid solubility and
toxic properties to parts of the DDT molecule (38 pp2990shy
2999) Rather they consider that toxicity is due to the
molecule as a whole and is explainable by means of a tr1shy
hedral1zed structure resulting from interaction of the
p-subst1tuted benzene rings and the bullCC13 group or groupshy
ings of similar and greater size
However a la relating toxicity to structure
broad enough to systematize and sympllfy the development
of successful agents needed t~ combat parasites afflicting
Man and his f~~d sources has not been forthcoming
The first step in any logical approach as to how
bigtlog1cally active substances w~rk is to record the
various biological physical and chemical properties of
active compounds~ Until enough relevant facts are colshy
lected progress toward such an end is slow and time
consuming Therefore it 1s hoped that the data g1ven
herein will add a small part to the necessary accumulation
ot evidence that may eventually settle the question of this
correlation or lack of correlation between chemical strucshy
ture and toxicity
The d1phenylmethylene radical Q bullCbull Q 1 was
chosen for this study of structural relationship to toxicshy
ity A review of the literature shows that this molecular
arrangement is contained in a number of biologically active
compounds
A preliminary investigation was first made in which
several derivatives of the basic structure were synthesized
in order to ascertain 1f certain functional groups ould
most readily lend themselvemiddota to study The results of th18
preliminary investigation indicated th t additional time
might pr~titably be spent contrasting the biological activshy
ity of ketone der-ivatives with the corresponding alcohols
Accordingly there were synthesized a series of
ketmes secondary alcohols and in the cases found feasible
the tertiary alcohols bull Chemical and biological data on
these compounds are given herein Other derivatives studied
in the preliminary investigation are als~ listed in an
addendum
The compounds were screened on mosquito larvae and
six microorganisms chosen for this purpose
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
THE BIOWGICAL ACTIVITY OF CERTAIN DERIVATIVES OF DDT
INTRODUCTION
It has long been recognized that the discovery of
a law rolating biological activity of organic compounds
to their structures would be of benefit to the investishy
gator who is unceasingly searching for agents exhibiting
such activity Selectively toxc agents in particular
are needed~ for agricultural foodstuffs must be protected
from weeds and pests of many kinds and economic animals
are troUbled with biological enemies of all types an
himself is n~t respected by these parasites
Up to this period of development in various fields
of the sciences ttempts have been made to correlate
chemical structure wit~ specific selective toxic activity
The insecticide DDT ll-b1s-(p-chlorophenyl)222-tribull
chloroethane m y be taken as a prime exam le for because
of its great economic importance many investigators have
given their time to considering this biologically active
compound
A first group considers the toxio portion of the
DDT molecule as res1d1ng 1n the linked p-ohlorobenzene
rings while the -cc13 grouping is believed to impart lipshy
oid solubility (22 pp892-928) Another group suggests
2
that the linked pbulloblorobenzene rings impart lipoid solushy
bility while toxioitr is due rather to the bility to split
out hydrogen chloride from the remainder of the molecule
(25 pp51~-513) A third group casts doubt on either of
these speculations and oonsidels that perhaps too simple a
relationship is sought tor such a complex phenomenon (6
ppl69-170)
A more theoretical approach has been taken recently
by yet anothe~ group who feels there is little basis for
assigning more or less independent lipoid solubility and
toxic properties to parts of the DDT molecule (38 pp2990shy
2999) Rather they consider that toxicity is due to the
molecule as a whole and is explainable by means of a tr1shy
hedral1zed structure resulting from interaction of the
p-subst1tuted benzene rings and the bullCC13 group or groupshy
ings of similar and greater size
However a la relating toxicity to structure
broad enough to systematize and sympllfy the development
of successful agents needed t~ combat parasites afflicting
Man and his f~~d sources has not been forthcoming
The first step in any logical approach as to how
bigtlog1cally active substances w~rk is to record the
various biological physical and chemical properties of
active compounds~ Until enough relevant facts are colshy
lected progress toward such an end is slow and time
consuming Therefore it 1s hoped that the data g1ven
herein will add a small part to the necessary accumulation
ot evidence that may eventually settle the question of this
correlation or lack of correlation between chemical strucshy
ture and toxicity
The d1phenylmethylene radical Q bullCbull Q 1 was
chosen for this study of structural relationship to toxicshy
ity A review of the literature shows that this molecular
arrangement is contained in a number of biologically active
compounds
A preliminary investigation was first made in which
several derivatives of the basic structure were synthesized
in order to ascertain 1f certain functional groups ould
most readily lend themselvemiddota to study The results of th18
preliminary investigation indicated th t additional time
might pr~titably be spent contrasting the biological activshy
ity of ketone der-ivatives with the corresponding alcohols
Accordingly there were synthesized a series of
ketmes secondary alcohols and in the cases found feasible
the tertiary alcohols bull Chemical and biological data on
these compounds are given herein Other derivatives studied
in the preliminary investigation are als~ listed in an
addendum
The compounds were screened on mosquito larvae and
six microorganisms chosen for this purpose
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
2
that the linked pbulloblorobenzene rings impart lipoid solushy
bility while toxioitr is due rather to the bility to split
out hydrogen chloride from the remainder of the molecule
(25 pp51~-513) A third group casts doubt on either of
these speculations and oonsidels that perhaps too simple a
relationship is sought tor such a complex phenomenon (6
ppl69-170)
A more theoretical approach has been taken recently
by yet anothe~ group who feels there is little basis for
assigning more or less independent lipoid solubility and
toxic properties to parts of the DDT molecule (38 pp2990shy
2999) Rather they consider that toxicity is due to the
molecule as a whole and is explainable by means of a tr1shy
hedral1zed structure resulting from interaction of the
p-subst1tuted benzene rings and the bullCC13 group or groupshy
ings of similar and greater size
However a la relating toxicity to structure
broad enough to systematize and sympllfy the development
of successful agents needed t~ combat parasites afflicting
Man and his f~~d sources has not been forthcoming
The first step in any logical approach as to how
bigtlog1cally active substances w~rk is to record the
various biological physical and chemical properties of
active compounds~ Until enough relevant facts are colshy
lected progress toward such an end is slow and time
consuming Therefore it 1s hoped that the data g1ven
herein will add a small part to the necessary accumulation
ot evidence that may eventually settle the question of this
correlation or lack of correlation between chemical strucshy
ture and toxicity
The d1phenylmethylene radical Q bullCbull Q 1 was
chosen for this study of structural relationship to toxicshy
ity A review of the literature shows that this molecular
arrangement is contained in a number of biologically active
compounds
A preliminary investigation was first made in which
several derivatives of the basic structure were synthesized
in order to ascertain 1f certain functional groups ould
most readily lend themselvemiddota to study The results of th18
preliminary investigation indicated th t additional time
might pr~titably be spent contrasting the biological activshy
ity of ketone der-ivatives with the corresponding alcohols
Accordingly there were synthesized a series of
ketmes secondary alcohols and in the cases found feasible
the tertiary alcohols bull Chemical and biological data on
these compounds are given herein Other derivatives studied
in the preliminary investigation are als~ listed in an
addendum
The compounds were screened on mosquito larvae and
six microorganisms chosen for this purpose
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
consuming Therefore it 1s hoped that the data g1ven
herein will add a small part to the necessary accumulation
ot evidence that may eventually settle the question of this
correlation or lack of correlation between chemical strucshy
ture and toxicity
The d1phenylmethylene radical Q bullCbull Q 1 was
chosen for this study of structural relationship to toxicshy
ity A review of the literature shows that this molecular
arrangement is contained in a number of biologically active
compounds
A preliminary investigation was first made in which
several derivatives of the basic structure were synthesized
in order to ascertain 1f certain functional groups ould
most readily lend themselvemiddota to study The results of th18
preliminary investigation indicated th t additional time
might pr~titably be spent contrasting the biological activshy
ity of ketone der-ivatives with the corresponding alcohols
Accordingly there were synthesized a series of
ketmes secondary alcohols and in the cases found feasible
the tertiary alcohols bull Chemical and biological data on
these compounds are given herein Other derivatives studied
in the preliminary investigation are als~ listed in an
addendum
The compounds were screened on mosquito larvae and
six microorganisms chosen for this purpose
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
4
Certain of the compounds under discuss on in th s
study have been examined from the viewpoint of biological
activity by other workers Following is given a brief
summary of the findings relevant to this investigation
Benzophenone has been extensively investigated as
a possible insecticide (33 pp985-991) and 24 pp43-47)
It is also known to be somewhat bocteriostat1o against
tubercle bacilli (15 ppl53-l56)
4 1 4 1 -Diohlorobenzophenone is kno n to be bacterioshy
static against tubercle bacilli (15 p154) and in addition
shows some insecticidal properties (27 p877)
Phenyl 2bullth1onyl ketone shows some toxic properties
toward the body louse egg (11 p33)
44bull -B1s bull(d1methylam1no)berlzophenone sho s some
slight bacteriostatic effect on tubercle bacilli (15 pl53shy
156)
Benzhydrol has been extensively investigated for
insecticidal properties and sho1s some act vity (36 p64)
and 10 p477)
41 4 1 -Dichlorobenzhydrol has aroused some interest
as an insecticide (27 p877) and 36 1 p64)
44-Dimothoxybenzhydr~l (36 p64) shows some
1nsoot1cidal activity against cockroaches and is known tgt
be bacteriostatic toward tubercle bacilli (15 pl53bull156)
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
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26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
5
1 lbullB1smiddot( p-chlorophenyl) ethanol has been dommshy
strated an active insect1o1de (16 pp361-362) and e
ppleS-111)bull It inhibits tnt growth or Cryptococcus ~shy
formanamp n_ vitro (40 p20) bull
B1sbull(pbullchlorophenyl)methane 1e someWhat to1o
toward mosquito larvae (9 middot ppllSll9 1nseets (27
p877) and rabbits (23 ppbull l27bulll42) bull
Some 1nseetic1dal activity has been demonstrated
tor llbullbla(p-chloropheoyl)ethylen$ 271 p B77_)
l 1lbullB1s-(p-cnlorophenyl)ethane snows toxic propbull
ert1es against 1nseets (44 p207) and 27 p877)
In summary then cllem1middotcE41 and biological data are
presented herein contrasting ketone and alcohol derivatives
of the d1phenylmetbylene rsltUcal
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
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Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
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ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
6
-------- - - - -middot--------- -----------
EXPERIMENTAL
Benzophenone
Benzophenone was obtained from the Eastman Kodak
Company Organic Sales D1vis1on~ Rochester 4 New York
Its melting point was found to be 47-48 degrees Centigrade
and was used with no further purification
44-Dichlorobenzophenone
44bullDiohlorobenzophenone waa prepared according
to the directions of Newton and Grogg1ns (34 pl39B)
Yields of 8207 per cent melting at 148 degrees Centigrade
were reported A yield of 78 per cent was obtained melting
at 148 degrees Centigrade
44-D1methoxybenzophenone
44-Dimethoxybenzophenone was obtained from the
Eastman Kodak Company Organic Sales Division Rochester 41
Ne r1 York Its melt1ng point was found to be 1435-145 deshy
grees Centigrade and was used without further purification
4a4 1 -Dichlorobull33bulldin1trobenzophenme
44-Dichloro-33 1 bulld1nitrobenzophenone was prepared
aoeord1ng to the directions of Backeberg and Marais (li
p805) A melting point of 131bull5 degrees Centigrade was
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
7
reported A yield or 746 per cent was found melting at
1320 degrees Centigrade
44-D1chlorobull33 1 -d1am1nobenzophenone
44middotD1chloro-33-d1am1nobenzophenone was prepared
according to the directions of Montagne (32~ ppl030-l031)
A melting point of 1675 degrees Centigrade was reported
A yield of 770 per cent was round melting at 1675 degrees
Centigrade
4bullChlorophenil ~-thienyl ketone
4bullCblorophenyl 2-thienyl ketone as prepared acshy
cording to the directions of Ng Ph BuubullHo et ~middot (7
pl095) A yield of 700 per cent melting at 99 degrees
Centigrade as reported A yield of 715 per cent melting
at 99 degrees Centigrade was found
Phenyl 2bulltb1enyl ketone middot
Phenyl 2bullth1enyl ketone was prepared according to
directions given in Organic Syntheses (4 pp520-52l) A
yield of 90 per cent melting at 55middot56 degrees Centigrade
was reported Ninety per cent yield melting at 55-56
degrees Centigrade was found
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
a 4 1 4-B1-s~(d1methylarn1no)benzophonone
44~middotB1s-(dimethylam1no)benzopbenone was obtained
from Eastman Kodak Company Organic Sales Division Rochester
4 New York Its melting point was found to be 173 degrees
Centigrade and was used w1 th no further purificatim
4-Fluoro-3-methylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 250 milliliter flask fitted 1th stirrer dropshy
ping funnel and reflux condenser were placed 200 grams
o-fluorotoluene 1n 150 milliliters dry carbon disulfide
To this were added 27 0 grams anhydrous aluminum chloride
Then 256 grams benzoyl chloride were slowly introduced by
means of the dropping runnel and the mixture finally reshy
fluxed three hours A condenser was attached to one of the
necks and the solvent removed The residue was poured on
100 grams of ice to which had been added 30 m111111ters conshy
centrated hydrochloric acid The mixture was extracted with
100 milliliters of ether washed once with 50 milliliters of
water once with 50 milliliters dilute sodium hydroxide and
twice more with water The solvent was removed and the
colorless 4bullfluoromiddot3-methylbenzophenone distilled at 176
degrees Centigrade under a pressure of ten millimeters of
mercury The liquid soon solidified and was twice crystslshy
l1zed from Skelly- aolve F to give 251 grams 645 per cent
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
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Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
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ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
9
middotyield) of the 4middotfluorobull3bullmethybenzgtphenone melting at 54
degrees Centigrade The calculated percentages of carbon
and hydrgtgen are 7849 and 519 found 7848 and 518 This
compound 1s identical in respect to melting pgt1nt and mixed
melting point to the 4-tluorobull3-methylbenzophenone given
below prepared by oxidation or 1bull(4-fluoro-3-methylphenyl)
lbullphenyletnylene obtained 1n turn from the ellucidated
structure ot 4-tluorobullS-methylacetophenone and phenylmagbull
nes1um bromide The 24-dlntrophenylhydrazone melts at
188bull189 degrees Centigrade Calculated percentages tor carshy
bon and hydrogen are 6091 and 383 found 6097 and 376~
Benzbydrol
Benzhydrol was prepared according to directions
given in Organic Syntheses (3bull pp90-9l) A yield of 69bull
72 per cent was reported melting at 68 degrees Centigrade
A yield of 70 per cent melting at 68 degrees Centigrade was
found
41 4middotD1chlorobenzhydrol
41 4-Diohlor~benzhydrol was prepared according to
directions given by Montagne (30 ppll5middot116) A melting
point of 94 degrees Centigrade was reported A yield of
85 per cent melting at 94 degrees Centigrade was found
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
10
4 bull 4 bullD1methoxzbenzhydrol
41 4-Dimethoxybenzhydrol was prepared according to
the method of Schnackenberg~middot (41 p655) A product
melting at 72 degrees Centigrade was reported A yield of
71 per cent m$lting at 72 degrees Centigrade as obtained
44bullD1chloro-33-d1n1trobenzhydrol
44-D1ohloro-33 1 middotd1n1trobenzhydrol was prepared
in the following manner In a 500 milliliter round-bottom
flask was placed a solution of 359 grams of pure aluminum
isopropoxide 1n 250 milliliters dry isopropyl alcohol and
200 grams of 44 1 -dichloro-3 1 3 1 -dinitrdbenzophenone A
short reflux condenser was attached to the flask but no
water run through the cooling jacket To the top of the
condenser was attached a water-cooled condenser set for disshy
tillation A boiling chip was added and the solution reshy
fluxed at such a rate that five to ten drops of distillate
were collected per minute After two hours when no more
acetone was found to be -present in the distillate water
was run through the upright condenser keeping the solution
under total reflux for fifteen minutes On retesting the
distillate with 01 per cent solution of 2 1 4-d1n1trophenylshy
hydraz1ne for acetone and obtaining a negative test~ the
excess isopropyl alcohol was removed under slightly
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
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Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
- - ---------------------~------------ -- -shy
ll
d1m1n1sned pressure The cooled mass we~ts hydrolyzed with
35 m1ll111t~rs of concentrated hydrochloric acid and 200
m111111ters watet_ An oil sep-arated whlob -as ampxtracted
with ether and washed with 100 milliliters water in two polbull
t1ons On removal of the solvent and after standing ove~
night the residue solidifiea was crystallized twice flom
bentene and thoroughly dried to give 182 grams of slightly
yellow 4 4--d1chlor~33bulld1ntrobenzhydJol melting at 110shy- shy
111 degrees Centigrade Calculated percentages for carbon
and hydrogen are 4550 and 235J found 4562 and 239 No
precipitate bullas obtained with 21 4bulld1nttrophenylhydraz1ne
under cond1tlons tbat gave a precipitate with the original
ketone
4 4 omiddotD1chlorobull33 bulld1am1nobenzhydrol was prepared
as follgtwst Two grams ot 44-dichlor~middot33-d1am1nobenzoshy
phenone vrere d1ssol~ed 1n l50 middotmtll111ters of anhydrous
methanol 1n a flask f1tted w1th a calcium chloride tube
F1ve tenths grams sodium borohydr1dmiddote were added and the
mixture was allowed to stand over n1ght at room temperature
The solution as t hen made strongly ac1d1e w1th dilute
hydroohlor1c ab1dt then strongly baste with dilute sodium
hydroxide Crystals separated when the sgtlution was poured
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
12
into twice 1ts volume of water and were f1ltered ana dried
One rectystall1zation from a benzene Skellybullsolve F milture
gave 181 graDl$ (90 per cent yield) of slightly yell)W
crystals or 4bull4 bulld1chloro-33middotdiam1nobenzhydrol melting
at 1185 degrees Centigrade as reported by Montagne (31
pp2260bull2261) for this compoundbull
4~0hl~rophenylmiddot2-th1enylmetbanol was prepared as
follows In a 100 milliliter round-bottom flask fitted
w1th a reflux condenser were placed f1 ve gramo of 4 ohloroshy
phenyl 2th1enyl ketone~ 60 m111111ters 95 per cent ethanol
10 milliliters concentrated ammonium hydrox1de a ten grams
of aluminum amalgatn The mixture was rerl~ecent1 six hours and
filtered while hot After cooling tf-e filtrate was poured
into twice its volume of watelbull Afte~middot standing 48 hours
crystals separatedbull On f1lter1ng 1 drying and recrystalbull
11z1ng from Skelly-solve B 3bull9 grams were obtained (77bull5
per cent yield) melting at 59middot60 degrees Centigrade as re
ported by Hamlin~~ bull for this compound (19bull p2732)bull
Phenrlbull2bullth1e-nzlmethanol
Phenylmiddot2bullthtenylmethanol was prepared by following
the procedure of Minnis 281 Pbull2144)bull A melting polnt ot
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
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Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
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Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
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Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
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40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
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Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
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ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
--
13
57bull58 degrees Centigrade was reported bull A yield of 79 per
cent was obtained mel t ing at 57bull58 de ees Cent ade
44middotB1s-(d1methylam1nobenzhydrol
44-Biebull(dimethylamino)benzhydrol was prepared
middot 1n the manner described by Yiastagl1 (26 pl657) 312
grams potassium metal were dissolved in ten mill111tera
dry benzyl alcohol Two grams of 44-Bls-(dimethylamlno)bull
benzophenone were added and the mixture was heated just
below reflux for t middot1o hours ~tater was added and the solshy
vent steam distilled The res~due was poured 1nto 500
m111111ters of water and after standing over night the
crystals were filtered and dried Crystallization from
Skelly-solve B gave 181 grams (90 per cent) of the hydrol
melting at 103 degrees Centigrade as reported by Mohla6
et al (29 p360) Mastagl i reported a 96 degrees Centmiddotbull
grade melting point
4-Fluoro-3-methylbenzhydrol
4bullFluoro-3bullmethylbenzhydrol was prepared as poundolbull
lows In a 100 m111111ter round-bottom flask fitted with
a mechanical stirrer were placed eight grams sodium hydroxshy
ide eight grams or 4-fluoro-3-methylbenzophenone 50 m1111shy
11ters of 95 pampr cent ethanol and eight grams zinc powder
The mixture was refluxed two hours 1 after which it was
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
14
filtered while still hot The residue was washed twice 1th
20 milliliter portions of not alcohol The filtrate was
neutralized 1th concentrated hydrochloric acid extracted
with 50 milliliters of ether and the ether layer washed
trice 1th 20 milliliter portions of ater The solvent
was removed and the hydrol distilled at 191 degrees Centishy
grade under a pressure of 14 millimeters of mercury to give
7 1 grams of colorless 4-fluoro- 3-methylbenzhydtol ( 66 per
cent yield) The percentages of carbon and hydrogen wer e
calculated to be 77 76 and 6 06 The percentages were
found to be 78 04 nd 6 03 A sample of the liquid gave
no precipitate 1th 2 4-d1n1trophenylhydra~1ne under conshy
ditions that a precipitate was obtained from the 4- fluoroshy
3-metbylbenzophenonebull
~ - ethylbenzhydrol
~-Methylbenzhydrol was prepared according to direcshy
tions found in Organ1e Syntheses (3 pp 226- 227) Yields
up to 75 per cent fere reported A yield of 69 per cent was
obtained after recrystallization from Skellybullsolve F melting
at 81 degrees Centigrade as reported by Klages (21 p 2646) bull
1 1- Bs (pbullchlorophenyl)ethanol
11-Bis - (pohlorophenyl) ethanol was prepared pecordshy
ing to dir ections given by Grummitt et al (17 1 p2265) bull
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
15
A yield of 89 per cent was reported melting at 67-68 deshy
grees Centigrade An 85 per cent yield as obtained meltshy
ing at 67bull 68 degrees Centigrade
l l -B1s-(p-methoxyphenyl)ethano1 was prepared
according to the method given by Grumm1tt l~middot (181
pp l269- l920) A yield of 83 per cent was reported meltshy
ing at 82- 83 5 degrees Centigrade A yield of 76 per cent
was obtained melting at 83 degrees Centigrade
1 - Phenrl- l ( 2~th1enyl) middotetllanol
lmiddotPhenylbulll2bull th1enyl)ethanol was prepared as folshy
lows In a 500 m111111ter three-neck flask were placed
646 grams or magnesium in 300 milliliters dry ether To
this was added slowly with stirring a total of 3 80 grams
of methyl iodide 1n 50 milliliters dry ether After the
magnesium had disappeared ~ total of five grams or phenyl-I
2- thienyl ketone 1n 50 milliliters dry ether was added with
cooling The mixture was refluxed one hour and poured on
50 grams ~f 1ce and 50 milliliters or water to which had
been added f1ve grams ammmium ohlorde The ether layer
was separated and washed twice with 25 milliliter portions
or water On removal of the solvent and recrystallization
from 95 per cent ethanol thebulle were obtained 402 grams
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
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Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
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26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
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Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
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34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
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Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
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Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
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ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
15
(74 per cent yield) of white crystalline middott-phenylbulll(2bull
thienyl)ethanol melting at 50 degrmiddotees Centigrade as reshy
ported b- Tb~mas (45 PPbull 643~644) ~middot
4bullFluoro3omethylaoetophonone was prepared in the
following manner In a 250 m1ll1liter thr~emiddotn~ck flask
f1 tted w1 th a mechanical stirrermiddot semiddotparatory funnel and
refllU condenser were placed ten gpams obullfluorotoluene 1n
150 mll11l1ters dry carbon disulfide To this were added
26 grams anhydrous aluminum chloride and the mixture heated
until gentle reflux began 9 27 grams ot acetic anhydride
were then added slowly over 15 minutes tlme Gentle reflux
was maintained tor two hours longer A condenser was then
fixed to one of the side necks and the solvent removed- The
contents of the flask were poured over 100 grams or 1ce to
which 20 mill111teJs concentrated hydrochloric acid bad been
added The mixture was extraoted twice wfth 30 m111111ter
portions Gt ethemiddotll atd the $ther washed twice with 30 m1ll1shy
11ter portions ot water once with 35 m1ll111ters 20 per
cent sodium hydroxide and then twice motte with 30 m1111bull
llter portione of water The solution was dried one houl
wlth five grams anhydrous calcium chloride and filtered
The solvent was then removed sndmiddot the 4bullfluorobull3-methylaoetobull
phenone distilled as a colotless liquid at 103 d6grees
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
17
Centlgr~de under a pressure f ten millimeters of merou~y
98 grams were bta1ned (7lG per dent yield) Percentages
of carbon and hydrogen were calculated to be 7104 and
5~96 found 7138 and 585 The 24bulldln1trophenylhydratone
melted at 232-5-2335 degrees Centigrade Calculated
percentages tor carbon and hydrogen were 5421 and 394
found 5402 and 3 86
It as deemed necessary to show that the structure
ot the 4bullfluol0bull3-methylacetophenone ts as given Accordbull
1ngly 200 milligrams of the compound were dissolved in
five milliliters dioxane plus one milliliter of ten per cent
sodium hydroxide Io~ine potass1tim~1odide solut1on made up
as advised by Shriner and Fuson (42~ pp138bull139) was added
with shaking until a slight excess yielded a dark color of
iodine at 60 degrees Centigrade After crystallization of
the iodgtform t~Tas complete the mixture was filtered deool3rbull
ized ith ten per oent sodium thiosulfate acidified with
d1lute hydrochlor1o acid_ and extracted with 30 milliliters
ether The ether layer was extractea with 30 m1ll111tere
ten per oent sodium hydrox1dmiddote which rtos ae1d1f1ed 1th
dilute hydrochloric ao1d to give 09 grams of white crystals
which when filtered and dried melted at 164-165 degrees
Centigrade given by Schiemann et al (39 p745) as the ~
mel tlng point opound 4bullfluorobull3-methylbenzo1o acid showing tbat
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
- - -~-~- ~--------
18
the aeylat1ng $gent attacks th$ pos1t1on para t o the
fluorine middot atom in o-fluoro-toluene undof thmiddote above
conditions
As turther proof of the strueture of 4fluorobull3bull
methyla o~tophenone 1 bull one gram middotport ton or 4-fluoro-3-methy-1shy
acetophengtne was placed 1n a 600 millilitermiddot threebullneemiddotk fl$Sk
11tted w1 th stirrer condensermiddot dropping funnel and heating
unit A saturated solution of potassium permanganate 1n
water was added tiVel a per1oamiddot 3f two hours until no more
reducmiddottton of the ox1d1z1ng agent took place wh1le under reshy
fltU The mixture was filtered and the filtrate acidified
with dilute hydttochloric acid causing white crystals to
separate These Were filtered and -r~dissgtlved in ZO millibull
liters of ten pet cent sodium hydttoxide ana reprecptated
with dilute hydroohloric ac1d t~ give 07 ~ams gtf white
crystals melting at 282bull266 degrees Centigrade bullmiddot as reporteel
by Fosdick --et al bull tor 4bullfluoro1sophthallo ac1a (141 p2309)
l-(4bullFluorn-3-methylphenyl)lbullpbenylethanol was pr e
parea as tallows In a 200 m111111ter thleebullneck tlamiddotsk
equipped with stirrer reflu~ condenser and dropping runnel
were placed 330 gtams magnesium 1n 50 m111111ters dry
ether To tttis were added slowly 217 grams bromobenzene
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
19
1n 25 mill111ters dry ether bull After the magnesium had disshy
appeared the flask as cooled and two grams of 4-fluorobull
3bullmethylacetophenone in 25 milliliters dry ether were added
slowly On refluxing for one hour t he contents ere
poured on 50 grams of ice to whtch 20 milliliters w ter
and f1ve grams ammgtnlmn chloride had been added middotThe ether
layer was washed twice with 20 milliliter portions of water
once with ten m111111ters of ten per cent sodium hydroxide
and twice more w1th 20 m111111ter portions of water On
evaporation of the solvent t he crystals were taken up in
Skelly-solve F and recrystal l ized to give 26 grams of white
1middot(4bullfluorobull3-metbylphenyl)l-phenylethanol (86 per cent
yield) melting at 63 degrees Centigrade Calculated pershy
centages for carbon and hydrogen are 7823 and 657 found
7801 and 649
lbull(4-Fluorobull3bullmethylphenyllbullph~nzlethylenamp
1middot(4-Fluoro-3-methylphenyl)lbullphenylethylene was
prepared as tollowst In a 250 m11l1litel tbreebullneck flask
fitted with stirrer dropping runnel and reflux condenser
were placed 640 grams of magnesium 111 50 m111111ters dry
ether To this were added slgtwly with stirring 424 bullms
bromobenzene in 50 milliliters dry ether After the magshy
nesium had disappeared the flask middotwas cooled and four grams
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
20
4-fluoro-3-methylacetopbenone were slowly added After
retluxing 15 hours the contents were poured on 50 grams
ice to which 20 milliliters of water and five grams ammoshy
nium chloride had been added The ether layer was sepashy
rated and washed t ice with 20 milliliters water once w1th
ten milliliters ten per cent sodium hydroxide and again
with t he same amount of ater The solvent was removed
two milliliters 20 per cent sulfuric acid were added and
the mixture refluxed one hour The organic layer was
separated and distilled at 172 de~ees Centigrade under
20 millimeters pressure of mercury to give 334 grams of
colorless liquid 1-(4-fluoro-3-methylphenyll-pheny1ethyshy
lene (602 per cent yield) Calculated percentages for
carbon and hydrogen are 8488 and 617 found 8499 and
516
4bullFluorobull3 ethylbenzophenone
4-Fluoro-3-methylbenzophenone was prepared as folshy
lows In a 25 m111111ter flask was placed one gram of
l-(4bullfluoro~3-methylphenyl)l-phenylethylene in ten millishy
liters glacial aoetic acid Two grams chromium trioxide
were added and the solution refluxed for one hour The
contents were then cooled and poured into 100 m111111ters
of water The crystals separating were filtered washed
well with water and taken up in ether The ether layer was
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
21
washed with 20 milliliters of ten per cent sodium hydroxide
and twice wi th 30 milliliters water The solvent was reshy
moved and t he solid recrystallized in Skelly-solve F to
give 750 milligrams of white crystalline 4bullfluoro-3-methylbull
benzophenone melting at 54 degrees Centigrade The calshy
culated percentages for carbon and hydrogen are 7849 and
519 found 7860 and 625
Compound screening on mosquito larvae
The insect repreeentative chosen for this study was
the southern house mosquito larvae Cul~x gu1n~ueasciatua
in the fourth instar It is hardy and adaptable to laborashy
tory conditions but perhaps less sensitive to toxic subshy
stances than other variet1e_s
The compounds were assayed in the following manner
Twenty milligrams of the compound were taken up in two
milliliters of acetone and 198 milliliters water By takshy
ing 015 and 15 milliliter al1quots and finally diluting
to 15 milliliters with ater the compounds were saayed at
levels of both one and ten parts per million
Round two-ounce bottles served as containers
Appropriate aliquots of sample were measured 1nto the
bottle and ten larvae 1ere added 1n five milliliters ~t
water The volume was then diluted to 15 m111111ters and
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
22
the bottlea allowed to stand The latvae v1ere aamined bullt
24 and 48 hour intervals the er1terion of death be1ng l ack
of respgtnse to probing
Results of tb1s screening atte listed ln Table II 1
block (g) as per cent of colony CJead after 24 hours applbull
cation of ten parts of egtmpouna per mllllm of water
The compounds under investigation ln this study are
quite water insoluble and it was found to be very difficult
to obtain rep~3duo1ble results in experiments utilizing the
photometer to assay bacter1ostat1e effects The convenshy
tional method of scrmiddoteenlng eompounds for bacter1ostat1c
propertieS using tihe optical density of an innoculated
culture media oonta1n1ng a known amount of compound for
estimating cell growth has other undesirable features
It is laborious time consuming and utilizes large amoubts
ot che~n 1cals when testing is attempted on a large scale
For these reasons a gradient plate technique
patterned aftel one reported by Bryson end Szybalsk1 (5
pp 45-5l) was investigated and dev-el~ped as a tool for
quick approximate Sereenlng for latge numbers of pottnshy
tial chemotherapeutics at a minimum of time and eGst-
Themiddot method is described as follows aseptic teohbull
nlque 1s used throughout till glassware being ster111zed
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
23
for 20 minutes at 15 pounds and t he nutrient a gar for ten
minutes at 15 pounds pressure To a plate nine centishy
meters in diameter placed at a slant as shown 1n Figure 1
are added 20 m111111ters sterile nutrient agar so that the
bottom is just covered
Figure 1
~I I After the agar has cooled and solidified the
plate is placed in t he normal horizont al position and
another 20 milliliter portion of sterile agar is added conshy
taining 200 micrograms penicillin G potassium salt See
Figure 2
Figure 2
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
24
Ttlamp thickness of agar 1s found to be about 063
centimeters After cooling two hours the agar plate 1$
lnnooulated by making a streak approximately 05 oent1bull
meters w1de bull1th a sterile brush dipped ln an 1nnoculum
Dpound stre2tococcua teoalis grown 48 hours in nutrient broth
at 32 degrees Centigrade The 1nnooulum streak follows a
llne bisecting the upper wedge of agar Finally the culbull
ture ta incubated 18 hours at 32 degrees It 1s then noted
that the colonies have grown ampmiddotlong the innoculum line beshy
g1nn1ng from the thin end of the top wedge ot agar as shown
1n Figure a
Flgule 3
Conatder a section of the agar plate along the lint
1nnoculated It 1a nine centimeters middotlong o63 centimeterbull
high and about os centimeters wide as shown 1n Figure 4
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
28
Plgure 4
liD I I I I I I I I I I I I I I I 1 z 3 ~ f J 10 n 1 13 1 1f I~ IT If
The aection may be divided into 18 smaller secshy
tiona each 05 oentimete~s in length with a volume ot 1575
oub1c centtmetera Section number one must contain appro21shy
mately 1-575 micrograms or pbulln1o1111n ott 10 m1orograma per
milliliter as this oonoentratlon was aCded 1n the top
wedge initially Due to downward cittus1on each succeashy
atve section will be deprived ot pen1clll1n that will be
estimated at one-e1ghteentb leas than the preceding section
By measurement it 1s determined that colonies ot
bull taeoal1s have atoppe4 growing at a poa1t1on dei1SQated
by aectlon number 16 Henoe it Ss calculated that the
colmies are inhibited by concentrations ot pen1cill1n of
about 17 micrograms per m1ll111te~ a figure within the
range found by other memiddotth)da (2 p72) Any oompoun4 may
be aesaye4 1n a s1m1lar manner
In evaluating the accuracy me m1gbt expect from
this assay lt is t~ be remembere~ that n~ 1ntegratl~n ~f
Clncentratlon along the gradient 1a attempted It 11
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
26
asawned that 41ftubull1on downward reaches equilibrium Out shy
ward diffust~n ts neglected The merit or this method
however 1s based )n the e~cellent reproducibility or reshy
sults and ~f the very rapid appr~~lmate $Yaluation ot large
nmnbers gtf potentially valuable compounds
The Yalues obtained by this method were compared
with those found by standard dilution technique Inooubull
lated tubes craquonta1n1ng ten mtll111ters of nutrient broth
and varying levels ot pen1c1111n G1 potassium aalt were
incubated at sa degrees Centigrade for 18 to 24 hours
The growth was then estimated by meana of a photometer
with reference to un1noculated tubes- The results are
tabulated 1n Table I middot
TABLE I
Smallest weight of pen1c1ll1n G potassium salt
Required to bompletely inhibit growth
Method
GJ~~gantam Gradient plate Serial dilution
1Ea cher1oh1a colt ___
~l tcrococcus nshyrosenese
13~0 )S bull
3
16 ~middot
ltl
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
27
A tot$1 ~t 32 compounds were then screened by tb~
gracUent plate meth~d and the ~eeults tabulated tn Teble
II~t Sis microorganisms wer chosen tor scresning rith middotbull
~1ew to obtaining ee varied a r~prcaentat1on ae teas1ble
All organisms w$ramp obtained feom stgtek cultures at oregotl
State College ~nd are as fomiddotllgtWll
(a
b)
Escherichia eol1 (sucrose n~gmt1ve)-Sta2hrlococcus aureus HSR9674t Horocoe~
(c)
(d)
(e)
(t)
mogenese
Streptococcus faecal1s ATT01170
Saechsromzces ell1Rao1deus (Burgundy wine)
Acetobsoter ~ubozdantJ ATCC621
Con141Q alb1cana
Tho values given 1n Table II are 1n micrograms
required to completely inhibit gJQwth The h1gbest level
taken for asetly was 250 miolograma per milliliter The
data 1e arranged tn the f~llowing mann$r
SAMPLE TABLE II
(a) c (d)(b)
-middot M s E s - taecil1s c011 RZ- el~JPbull usName ot sbullnes Strua
pnnpound ture (e) (g)t)A Culexc
aubogshy - gu1nguefasc1atuaalbicansCJane
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
28
TABLE II
- -Benzophenone o-~-o
n~t - -run
(14 112 bullOH
Benzhydrol o-~-o - not run
bullbullbull bull-OHO ~-methylbenzhydrol O-J I
- not -run
Q
4 4 bulld1chlorobenzobull phenone o-C-o01 Cl
not -- run
- 112 140 42 OH O~ -c- I44-d1chlorobenzshy
hydrol ClO H ampcl 140
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
29 TABLE II (Continued)
11-b18-(p-chlorobull phenyl)ethanol r- - - ~elOcs 01
~ 42 42 42
36
44 1 -d1methoYbenzoshyphenone
o-La~H- b oms
- - - -
- not run -
44-d1methoxybenzbullhydrol J8-~~a
~H H Ac
- - -
- not run -
11-bia-(p~metbo~y-phenyl)ethanol r)cQ
~II ~~
- - -
- not run -
44-dlcbloro-33shyd1n1trobenzophenone c1 o--oI ca
NOe N02
- - -
- -
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
TABLE II (Continued)
44-d1ohlorobull31 3tbull dinltrobenzhydrol
0 OHO-o- I ~ Cl H Cl
N02 NOt
14 70 - lt14
lt14 not run bull
44bull-dlchloro-33shyd1aminobenzophen~ne
oJ-oc1 c1
NHa NB2
- -
middot not r~
-
4 4bull~1ohlorobull33bull d1am1nobenh-rol
OHo-a-oCl H 01
Ime NHe
- - -
- not run
4-chlorophenyl ashyth1enyl ketone o-tuCl
bull
not run
31C
4bullChloropheny1middot2shyth1enylmethanol
OH
Clo-~middotu 70 112 70 42
42 - bull
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
TABLE II (Continued) 51
oJ-u - - bull 54
phenyl 2bullth1enylketor1e
- not bull run
182 - -OH O phenyl-2-th1enylbull o-~ -methanol
- ngtt ~
run
o-centaLJ ~24 70 l-phenyl-1bull(2shyth1enyl) ethanshyol CH3shy
- ngtt -run
0 gt bull
4r4 bullb-iabull( d1methyl o-~-aamno)ben~ophenone li I
( OH3) 2 ( CH3~ - not run
f
o OH a - - 42
44bullmiddotb1s-(d1methylshy -c I amino) benzhydrol W H
I I lt14 not (OH3)2 (CH3~ -run
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
TABLE I I (Continued )
- - middot- 0
4bullfluraquorobull3-methylbull o-amp-obenzophenone F
3 not -run
OR lt14 lt14 lt14
4bulltluoromiddot3~ethylbull o-c-obenilhydrol F H
CH3 lt14 not -run
- 42 lt14 lt14 1bull(4bullfluoro-~middot - bull ~ meth1lpben7l)lbull Po 3ophenylethanol
OH3 - -13 3 1bull7
pen1o1ll1n Ga potas sium salt - not
run
90 650 -
phenol
- not not run run
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
33
DISCUSSIOW
In reviewing the data given 1n Table II regarding
screening ~f the series of compounds- on microorganisms bull
~ne fact is immediately apparent or the four types of
bacteria studied not gtne was inhibited by a ketone
Phenyl 2bullthienyl ketone in c~ncentrations of 54 microshy
grams per m111111ter inhibited the grwth of the yeast
[bull ell1pso1deua however Apparently ket~nea ot tb1a
type are relatively innocuous to the organisma studied
In contrast to the ket~nes reduction to the
alcohol derivatives in general yielded highly toxic m~leshy
oules (f the nine ketonesmiddot assayed seven when reduc-ed
gave alc~hols proving toxio in ooncentrati~ne varying from
less than 14 t~ ab~ut 225 micrograms per milliliter Only
the methosybull and aminobullchloro substituted derivatives
tailed to show toxic pr~pert1es
The unsubstituted benzhydrol 1nh1b1ted the growth
of both bull 22 and _ 27l06en~se 1 but those alcohols w1 th
additional substitution tended to prove toxic t~ a wider
spectrum ~f organisms
The ~-methyl substituted benzhydrole ah~wed varied
effects For benzhydr~l itself toxicity was eliminated
For 41 4 1 -d1chlor~benzhydrol alth~ugh toxicity was remoted
tor the pathogenic yeast two microorganisms bullere inhibited
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
34
to a greater extent lhe 4bullfluorobull8-methylbenzhydrol was
made more acceptable by the addition of an o(-methyl gl~up
but ln contrast the toJC1c1ty opound phenylbull2bullth1enyl methanol
was increased
The tluoroillethyl substituted alcohols showed b1gb
toxicity exceeding that exh1b1ted by the chloro der1vabull
tivea Addition ot nitro groups to aid chlorine 1llustrate4
by 441-dichlorobull$ 1 3-dlnittobenzbrdrol gave a molecule
quite efteotive 1n hindering glowth
The thiophene rtng substituted for phenyl gave a
somewhat wider apectztum of aot1v1ty although tn several
cases the compound was somewhat less toxio
Although an amino group in the 3 position aa
illustratedmiddot by 44 t -dichloro-33 -d1am1nobenzhydrol was
innocuous the tertiary amino group in 44middotmiddotb1s-(d1methylshy
am1no)benzhydrol yielded a compound qu1te effective tJr bull ell1p~1deus and bull Suboxzdans
bull ezrogenese end bull ell1paoldeua proved most susshy
ceptible both belng inhibited by e1ght compounds c -
alb1cena as most resistant being affected by only 41 4tbull
diehlorobenzhydrol This is to be noted 1 hJwever bull tgtr tha
pathogeni-c yeast is extremelr resistant to most chemotherashy
peutic agents
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
Of the c~mp~unds listed in thbull addendum llshy
bls(4bullchl~ro-35-dinitrophenyl)ethane proved t~ be quite
to-ic as did the bis-(4-chlorophenyllacet1c acid
Only three oompounds proved to inhibit insects
to any extent the 4-chlorophenyl 2bullthienyl ketone alone
being investigated in this capacity for the first time
Future investigat1gtn in this direction might be
well spent determining the toxicity or these compounds
on higher animals If favorable results are obtained
further research pertaining to feasible alcohol derivashy
tives or this general type might furnish valuable apecltlc
toxic agents ot practical importance
The gradient plate method for rapid screening of
pr~speotive chem~therapeut1c agents shows great promise
Further development ~f this meth~d would certainly be
profitable
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
36
(
P~llowing is a brief aum~ry ot this work
1 A rapid gradient plate method ~f screening
potential -chemotherapeutic agents tbull repotbulldbull
a Thirtybulltwo compounds have been screened on four
bacteria two yeasts and one insect
~ There te a definite contrast 1n the b1olog1cal
activity of the ketones and alcohols prepared the former
being nonto21c ln the concentrations used
bullbull Si2 new compounds plus derivatives are among
those reported
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
s
5
a
10
11
12
37
BIBLIOGRAPHY
Backeberg o G and J L o Marala Some ~er1vat1ves ~~ lallbulltr1ohlorobull2a2-d1bull(4bullohlorobullphenyl) ethane (DM) bull Journal of the obem1oal society 803bull806 1945
Beyer Xarl H Phatmacolog1cal basta ot pen1c1111n therapy Spr1ngfleldbull Thomas 1950 214p
Blatt_ A H (ed) Organic fiyntheaea Colleot1vbull vol 1 ld ed- Nbullbull York Wlleybull 1941 680p
Blatt A H (ed) Grganic syntheses Collective vol 2 ftew Yorkbull W1leyt 1943 654p
Bryaon middotvernon and waclaw Szybalsk1t 1crob1al eelecshytion Science 11645-51 1952
Buav1ne1 J R middot Insect1c1del action ot DDT Nature l66t~69middot170 1945
BuubullBo NgPh Ng Hoanand Wg D Xuong ~tudes dans la aerie du thiophene v N)uvelle cetone thiopniques et leara deriva Recue11 debull traYaux oh1m1ques des payabullbas 69tl083middot1108bull
Dean R w Summer C3ntrol of orchard m1tee 1n eaatshyern New middotYork Journal of economic entomology 43t 167bull171 1950
Deonier Christian c et al Soap and sanitary chem1shycala 22bull Number lltl11J-Ir9 1946 (Abstracted ln Chemical abatracta 4l(l)ti50t 1947)
Eaton1 J K and R G Daviea The toxicity ot o6rbull ta1n 8ynthet1o organ1c compounds to the tru1t-tlee red-apider mite Annals f applied biology 37c47lbull 489 1950
Eddy Gbull1na w and N B Carson Organic oompoundatested a gainst body louse eggs Journal opound economi~ entomology 4131bull36 l94B
Fischer~ E t middot C Garces bull and A Lopez Relation bebull tween qu1no1d structure and bacter1ostat1c activity ot tetramethyl-d1am1nod1phenylmethane derivatives Journal of bacteriology 5lal-8 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
38
17
18
21
22
Forrest Jamea Oliver Stephenson and 1111am A aters Chemical invest1gat1one of the 1naect1c1de
DDT and tts analogues Part 1 Reactionbull of DDT and associated cgtmpounds Tournal or the chem1oal aocletr 333bull339bull 1946bull
Fosdick L s and J o Calandra Synthea1a of blsbull ( d1a1kylamtnoalkyl) esters of 4bullfluorotsophtal1oaold lournal of tne American chemical society 65t2308bull2309 1943
Preedlander~ B Lbull Effect of benzophenone and allied oompounds on human tubercle bacilli in vitro Pr)oeetUnga of the s ooietr tor esperiiental middot b1olOSJ and medicine 61153bull156 19~
Grummitt Oliver BSbull(p ohlolophenyl)methyloarbnol 1 a new m1t1c1demiddotbull Science lll 361bull362 l960w
Grummltt Oliver Allen E Buck and Ernest I Becker 1lbullD1bull(p~ehlorophenyl)ethane Journal of the American ohem1cal society 672265bull2266 1945
Orummttt I 011ver and Dean Marsh D1-p-bulln1ayl)methyloarbtnol Journal of the American chemical society 701289bull1290 1948
Hbullmlin K Ebullbull et al 4~hlorophenylmiddot2bulltblenylbullmethanol Journal ot the American chem1oal aociet7 71231bull2734 1949
Dng1 Harold 0rtho-parabull1Somer1sm 1n the prbullparat1on or d1am1no41phenylmethane Journal or the chem1cal aociety ll7a96Bbull992 1920
Xlagea August Syntheae dia~lrter AethyleneBer1chte der deutbullohen chemlacben Geaellachaft 3526middot6~2649 1902
Lauger P- H Martin and P Mflller ~bullr Konatitubull tlon und to21ache 1rkung von naturl1chen und neuen synthetischen 1naektent6tenden Stotfen Helvetica ch1m1ca aota 27892bull928 1944
L1111e bull R D lA I Sm1th ana E F Stoblman Pathoshylogic action ot DDT and certain of ita analogs and der1vat1vea Archives of pathology ~127bull142 1947 (AbstracteltJ in Chemical abstracts 41(2)bull 6967b 1947)
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
26
28
31
32
L1nduska J P F A Morton and ~ c McDuff1eshyTests or materials ror the control of chiggers on the ground Journal of econnmic entomology 4lt 43bull47 1949
Martin Hbull and R L ~ ain Insecticidal action of DDTbull Nature l54t512-513~ 1944
Jlastagll Plerre Aoton r6ductr1oe et condensatr1oe des benzylate$ aleallne sur les ctones et lee aldhyclee et sur lea alcools el i non satUJeebull Comptes rendus des seanoes de lbullacademie des ac1encea 204sl656bulll656 1937
Metcelt Robert L Acaricidal properties of organiccompounds related to Din Journal of eemom1c entomology 4ltS75bull882 1948
Minnie Wesley Some th1opbene analogs of dibull tr1bull and tetraphenylmethane compounds Journal of the American chemical society 5lt2143bull2144 1929bull
M8hlau Richard and M Heinze Synthesen aryl1rter Leukaurbullm1ne~ Ber1chte der deutschen chem1schen Gesellsohatt 35t358bull375 1902-
Montagne v P bull J Sur les transpositions atomiquea1ntramoleeula1rea Recue11 des travaux oh1m1queades pays-bas 24amp105-131 1905
Montagne P J t1ber die E1nw1rkung alk~holiseher Kal1lauge aut Halogen-aminobenzophenone(undbullbenzhydrole Ber1chte der deutsehen chemtschen Gesellschaft 492243middot2262 1916
Montagne P J Uber die Konstitution der durch Nttrlerung dea 41 4 1bullD1chlor-und des -amp4-Dichlorbull d1n1tro-und 414iD1brom-d1n1tro-benzophenoneBerichte der deutschen chemischen Gesellaohaft 481027-1037 1915~
Nagase lakoto Insecticidal principle ln the smoke produced by burning insecticide powder Journal ot the agricultural chemical society of Japan 17t986-991 1941 (Abstracted 1n Chemical abstracta 452140d 1951)
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
36
40
34bull
35
38
40
Newton ~middot r ppd P H Groggins Benzophenonea tom carboYl1c acids Industrial and engineerbull1ng chemis~ry 271397bull1399 1935
Pfeiffermiddot Rbullmiddot amiddotnd R W1z1nger zur Theorie der Halogenaubat11on Annalen ~er ch1m1e 461132bull 154~ 1928
Picard~ Tean P tlnd c w Kearns Analysis of the essential structural features of DDT by a study of the tox1o~ty of closely related cnnpounds to roaches and to housetl1es Cana~1on journal of research 270t59bull67 1949
Price Charles c and George P Mueller The p1nashyools and middotp1nacolones rrom pDethoxyacetophenme bull Journal ot the American chemical society 66t634bull 636 1944~
Rogers E F et al The structur~ and toxicity of DDT 1nseet1oYOeS Journal of the Amer can ohm1cal e~c1ety 752990middot2999 l95S
Schiemann Gftnt~er and Wilhelm Roael1us Uber aromatische Fluorverbin~hmgen bull X M1tte1lt HShershyflu~r1erte D1phenylverg1ndunger Ber1chte der deutschen chem1sohen Gesellscbatt 65737bull745 1932
Schmidt Bm11 a bullbull et ~middot Amicr~b1~l~g1oal study of Crzftpcoooua neOrormana Archives ot biochemistry16t -Si 1915obull
Schnackenberg Hans and R Scholl Uber daa p-Dibull meth~xybenzhydrol Ber1ohte der deutaohen ohem1sch~n Oesellschatt ~6r654-655 1903
Shrinerbull Ralph L and Reynald c Fuson The systemshyatic 1dentlt1cat1on of organ1o compoundamp 3d ed Bew York 11ley 1948 370p
Sra1th1 M~ I et al The pharmluological action ot certa1n ansigUis and derivatives of DDT J~urnal ot pharmacology and e~per1mental therapeutics 88 359-365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
Stringer A The 1nsect1c1dal act1m of some DDT tanalogues and chlorinated (4-cblorophenyl)shyeth4tnea Annals of applied biology 36s206bull212 1949
Thoma a M v Sur quelquesdet1vea dee th1opene bull
Comptes rend us de 1 bullacademe des ac1enoee 14Eh 6~middot648 1908
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
ADDENDOX
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
--
ADDENDUM
B1a~(pbullohlor~phenyl)metb~n$ wss prepared acoo~d1ng
to lt11reet1ons given by Smith et al (5 p364) A heobull
ret1cal yield melting at 55bull56 degrees Centigrade was reshy
ported A neal theoretical yield melting at 55bull56 degrees
Centigrade was obtained
~1lbullBis-(pbullchlorophenyl)ethylene
llbullB1sbull(pbullohloropheny-l)ethylene was preparelt bJ
the metbod glven by Grumm1tt bull (2~ pp2265bull2266) A
yield of 88 per cent melting at 8~middot86 degrees Centigrade
was reported A yield of 68 per cent melting at 84bull86
degrees Oent1grade was found
llbullB1abull(p-methoxyphenyl)etbylene was prepared
acco~d1ng ~o the method given b Pfeiffer and W1~1nger
(3 pl44)- A y1eld ot 74 per cent bullas reported melting
at 1-2bull143 degrees Centigrade A yield of 195 per eent
meltlng at 142-143 degreea Centigrade was obtained
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
llbullB18bull(pd1methylam1n)phenyl)ethllene
ll-Bisbull(p-d1methylam1nophenyl)etbylene na preshy
pared acc~r4ing to directions given by Pfe1fer and
W1z1nger (3 pl52) A 56 per cent yield ns reported
malting at 121bull122 degrees Centigrade A 59 per cent yield
was foulld melting at 121bull122 degrees Centigrade
ll~B1s-(pbullohlorophenyl)ethane was prepared accordbull
lng to the method or Grumm1tt bull lte p2266) A
yield of 62 per cent melting at 54-55 degrees Centigrade
was reported A yield or 63 per cent melting at 54 55 deshy
grees Ce t1grade was found
llbullBlsbull(p-metho~yphenyl)e~hane was prepared aa
tollowas a m1zture or ao grams l lbullb1sbull(pwmetbozypbenyl)shy
bullthylene 200 mllllliters absolute alcohol and eoo milllshy
grame copperbullchrom1te catalyst was allowed to react with
bydrogen ln a bmlb at 175 degrees Centigrade and 2000 pounds
pressure Attel two hours the bomb was removed The eolubull
tion was filtered h~t the a~l~ent rem~velaquot The product
bullbulls crystallized fr)m 95 per cent bulllcohol to give 13 srbullmbull
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
--
44
(65 per cent yield) of ltlbullbis (p-methoxyphenyl)ethane as
reported by Price and ueller tor this compounct (4 p636)
llbullB1ebull(bullmiddotchlorobull3Qbulld1n1tropnenyl)ethbullnbull waa
prepared according to the method given by Forrest 1 lbull (1 p33B) A product melting at 261 degrees Centigrade
was reported A yield of 52 per cent melting at 261 dbullmiddot
greea Cen~igrade was toand
Bia-( pbullahl oropbenyl) acetic acid
B1s-tpbullchlorophenyl)acet1c acid was prepared aooordshy
ng to directions g1lVen by Smith et al (5 pp364-365)
The~e was reported a 353 per cent yield which melted at
1675bull168 degrees Centigrade There was found a 396 per
cent yield which after o~ystall1sat1on from 36 per cent
acetic acid melted at 164 degrees Centigrade
Compound aoreen1ng
The above compounds were treated tor acreenins
exactly aa berore and tt_le results are 11ste4 ln Table III
preotael7 as 1n Table II_
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
45
TABtE III
- - -0 CH3011-bisbull(pbulloblorgtbull o1 -~- I 1phenyl) ethane not 5~run
o-ra - -llbullbia-(plethozrbull
phenyl) ethane Pf5 H ~ Clt5 pegtt -un
CH30 - ~12 lt14 lt14
11-bismiddot(~chloro- ~o-~- I Cgtz35-d1n1trophenyl)ethane Olshy 1
NOg N0middot2 112 0 - lt14 lt14
o~~Ho~iabull(p-oblorophenyl) Cl - Hshy I ~ lacetic acid po
run
CHg - - - 1middot(4-fluoro-3bull o-C-o~ethylphenyl) lbull F I jPhenyletbylene
CH - pot 3 run
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
TABLE III (Omt1nued)
-t
bull - --c- Ib11bull pbullchloropheny1) OHO methane Cl H b el not- run
- --l lbullbia-(p-chlo~omiddot o-r-ao
Cl 1pbenyl)ethylene not bull run
bull bull-CH2 middotshy1 1-bia-(pbullmeth~xy- o-~- aplleny1)ethy1ene ~ -ltXE~ not- -run
- - --~1middotb1s-(p~d1methylbull
am1nophen)l) o~ruethylene ~CH3)2 (OH3)2 ~ot -
ltun
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946
4ll
ADDENDUM
BIBLIOGRAPHY
1 Ps_rreet James bull Oliver Stephenson end William A Waters Chem1oal 1nvest1gat1ona or the inaectlmiddotctde DDT and lts analogues Part 1 Rbullactlone ot DM and aseoc1ated compounds Journal ot the che~ical society 333bull339 1946
2 Grumm1tt Oliver Allemiddotn E Buck and ~rnest I Becker 11-Dl~pbullcblot~phonyl)ethane Journal of the American chemical society 67amp2265middot2268 1945
3 Ptelffer R 1 and R W1 z1nger Zur Theor1emiddot der Hal~genaubst1t1on Annalen ~er ch1me 461132bull 154 1929
4a Price Charles c and Gegtrge P Mueller The p1nashyoo1a middot and p1naoolones tr~ pbullmetho~yacetophenone Journal of the American chemical S3c1ety 66s6254bull 636 1944
5 Smith bull I bull - et al The pharmacological aotl~n ot certain anirogues and derivatives ot DDT Journal of pharmacology and experimental therapeutics 88 359bull365 1946